In vivo, real-time confocal imaging

J. V. Jester, P. M. Andrews, Walter M Petroll, M. A. Lemp, Harrison D Cavanagh

Research output: Contribution to journalArticle

94 Citations (Scopus)

Abstract

We have adapted a tandem scanning confocal microscope for real-time, non-invasive imaging of cells under in vivo conditions. This form of in vivo confocal imaging relies on the optical sectioning abilities of the confocal microscope to obtain en face, sequential, reflected light images of cells at various depths, up to 1 mm, within opaque organs in living animals. Of major consideration in the design of an in vivo confocal microscope is maximizing the real-time detection of signals reflected from low contrast structures which can be affected by the microscope design, objective, and image detector systems. Using an in vivo confocal microscope design with a 20x BioOptics surface contact objective we have obtained live cellular images from selected tissues including cornea, kidney, liver, adrenal, thyroid, epididymis, and muscle and connective tissue of rabbits and rats. Images were captured, digitized, and processed using a DAGE Mti low light level SIT camera coupled to a Gould IP9527 image processor. In vivo images were also compared with conventional bright field light and scanning electron microscopic images of 'dead,' fixed tissues. Overall, in vivo confocal imaging can provide remarkable detail of living cells comparable to that of conventional microscopic images of 'dead,' fixed, and stained tissue. A more unique feature of in vivo confocal imaging is the ability to study cellular structure and function sequentially over time in the same organ or tissue and represents a fundamentally new paradigm in microscopy. With continued refinements in the microscope, objective and detection system designs and their consequent improvements in lateral and axial resolution, in vivo confocal microscopy will enable us as observers to see what no one has been able to see before.

Original languageEnglish (US)
Pages (from-to)50-60
Number of pages11
JournalJournal of Electron Microscopy Technique
Volume18
Issue number1
StatePublished - 1991

Fingerprint

Microscopes
Imaging techniques
Tissue
microscopes
Light
Epididymis
Cellular Structures
Confocal Microscopy
organs
Connective Tissue
Cornea
Scanning
Microscopy
Thyroid Gland
Confocal microscopy
Electrons
Rabbits
microscopy
Kidney
connective tissue

Keywords

  • Confocal microscopy
  • Light microscopy
  • Scanning electron microscopy

ASJC Scopus subject areas

  • Anatomy

Cite this

In vivo, real-time confocal imaging. / Jester, J. V.; Andrews, P. M.; Petroll, Walter M; Lemp, M. A.; Cavanagh, Harrison D.

In: Journal of Electron Microscopy Technique, Vol. 18, No. 1, 1991, p. 50-60.

Research output: Contribution to journalArticle

Jester, JV, Andrews, PM, Petroll, WM, Lemp, MA & Cavanagh, HD 1991, 'In vivo, real-time confocal imaging', Journal of Electron Microscopy Technique, vol. 18, no. 1, pp. 50-60.
Jester, J. V. ; Andrews, P. M. ; Petroll, Walter M ; Lemp, M. A. ; Cavanagh, Harrison D. / In vivo, real-time confocal imaging. In: Journal of Electron Microscopy Technique. 1991 ; Vol. 18, No. 1. pp. 50-60.
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